Variable repetition rate and wavelength optical pulse source

ABSTRACT

Variable repetition rate and wavelength optical pulse source, comprising a fixed or variable repetition rate source of supercontinuum pulses; a wavelength tunable optical bandpass filter to filter the supercontinuum pulses at two or more wavelengths, wherein said source of supercontinuum pulses and said wavelength tunable optical bandpass filter are configured such that the optical pulse source can provide variable repetition rate and variable wavelength optical pulses including a series of repetition rates with selected wavelength-varying pulse trains.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.12/316,006, filed on Dec. 9, 2008, which claims the benefit of priorityunder 35 U.S.C. §119 of Great Britain (GB) Patent Application Serial No.0800936.7, filed in the United Kingdom on Jan. 19, 2008, whichapplications are hereby incorporated by reference herein.

FIELD OF THE INVENTION

The present invention relates to optical pulse sources and methods ofusing, such as, for example, as an illumination source for a StimulatedEmission Depletion (STED) microscope including the optical pulse source.

Supercontinuum lasers delivering continuous spectra over an ultra-broadbandwidth offer an ideal source for many applications within biomedicalimaging, component characterisation, manufacturing control and defenseresearch. The most common form of supercontinuum laser comprises anultrashort pulse optical pump source operating at a pump wavelength inthe Infra-Red (IR) region of the spectrum (typically around 800 nm for aTi:Sapphire laser or 1064 nm for mode-locked fiber lasers and diodepumped solid-state lasers) and a highly nonlinear photonic crystalfiber, with specially designed dispersion properties. The interactionbetween the high intensity optical pump pulse and the nonlinear silicafiber causes extreme broadening into the visible and infra red regionsof the spectrum to provide spectra spanning from approximately 450nanometers (nm) to beyond 2.5 micrometer (μm).

The phenomenon of supercontinuum was first proposed in the 1970's byAlfano et al (see R. R. Alfano and S. L. Shapiro, “Observation ofself-phase modulation and small-scale filaments in crystals andglasses”, Phys. Rev. Lett. 24, 592 (1970)) and in 2000 the firstdemonstration of a fiber-based supercontinuum made by Ranka and Windelar(see J. K. Ranka et al., “Visible continuum generation in air-silicamicrostructure optical fibers with anomalous dispersion at 800 nm”, Opt.Lett. 25 (1), 25 (2000); see also U.S. Pat. No. 4,450,427) using aTi:Sapphire mode-locked femtosecond laser 10 to pump a photonic crystalfiber 11 with zero dispersion wavelength at approximately 760 nm in freespace through launch optics 12 as shown in FIG. 1. More recently,researchers and companies have utilized nanosecond and picosecond-basedpump sources operable at around 1064 nm wavelength to pump photoniccrystal fibers with a zero dispersion wavelength close to 1 μm togenerate high brightness supercontinuum products.

Commercial supercontinuum products, such as Fianium Limited'ssupercontinuum fiber laser model no. SC450, rely on mode-locked fiberoscillators and high-power optical fiber amplifiers to generate highlyintense pulses at the pump wavelength, which when injected into anonlinear photonic crystal fiber (PCF), result in extreme spectralbroadening into both the visible and IR regions of the spectrum.

FIG. 2 shows an example of such a system, where a mode-locked fiberoscillator 21 produces low energy optical pulses of approximately 10picoseconds (ps) duration and at a repetition rate of 20 MHz to 100 MHz.The pulses are amplified within a cascaded fiber amplifier 22,comprising one or more stages of amplification, with optical isolation23 between each stage. The output of the amplifier is a high-energypulse of up to 400 nJ and tens of kilowatt peak power. The output of theamplifier is injected into a length of highly nonlinear photonic crystalfiber (PCF) 24 with anomalous dispersion at the pump wavelength (1064nm) and zero dispersion at a wavelength close to the pump wavelength.The spectral bandwidth of the pulse broadens within the PCF, creating apulse of approximately 100 nJ energy and a bandwidth from 400 nm to 2.5μm that exits the PCF 24.

The pulsed nature of these sources and high repetition rates (20 MHz to160 MHz), makes these sources attractive to both applications requiringquasi-continuous wave radiation as well as applications involvingtime-resolved measurements such as FLIM (Fluorescent lifetime imaging)and TCSPC (time correlated single photon counting). However, often, thehigh repetition rates can be a limiting factor in lifetime imaging,where the pulse-to-pulse separation (50 ns for a 20 MHz source, down toa few nanoseconds for a 160 MHz source) can be much shorter than thelifetime of the sample under evaluation. In this event, one requires alower pulse repetition rate.

In general, most mode-locked laser systems operate at pulse repetitionrates of several tens of MHz (diode-pumped solid-state (DPSS) laserstypically operate from 80 MHz to 100 MHz, and fiber lasers from 20 MHzto several hundreds of MHz). Some applications require lower repetitionrates—for example fluorescent-imaging lifetime microscopy (FLIM), wherefluorescent-labeled molecules are excited by an incident optical pulseand the decay of the fluorescence is monitored before the next opticalpulse arrives. In FLIM, the lifetimes of interest can very often exceedtens or even hundreds of nanoseconds, and therefore it is often requiredto have repetition rates of 40 MHz and below. Repetition rates of lessthan 20 MHz, while not impossible to achieve, are difficult to deliverfrom a mode-locked fiber oscillator due to high nonlinearity within along cavity (10 m for a 10 MHz oscillator). From a DPSS source, such acavity (5 m long for 20 MHz and 100 m in length for 1 MHz) is almostimpossible to make due to the required complexity of the cavity design.

SUMMARY

According to a first aspect of the invention there is provided anoptical pulse source comprising an optical pump laser operable togenerate optical pump pulses at a pump pulse repetition rate. The sourcealso includes a nonlinear optical element, and a gating device providedbetween the pump laser output and the nonlinear optical element. Thegating device is operable to selectively control the launch of pumppulses into the nonlinear optical element at a variable, lowerrepetition rate, to thereby selectively control the generation ofoptical supercontinuum pulses within the nonlinear optical element andtheir repetition rate.

The optical pulse source enables optical supercontiuum pulses to begenerated with flexible control of the output pulse repetition rate, andthus the pulse-to-pulse separation. Control of the inter-pulseseparation is of particular advantage where the source is used toprovide supercontinuum illumination in applications where decaylifetimes are measured, such as FLIM, and the optical pulse source canbe operated to generate supercontiuum pulses at pulse repetition ratesof less than 20 MHz for such applications.

The nonlinear optical element may comprise one or more nonlinear opticalcrystals, such as lithium triborate (LBO), barium borate (BBO), cesiumlithium borate (CLBO), periodically poled lithium niobate (PPLN) orperiodically poled potassium titanyl phosphate (PPKTP) or amicrostructured optical fiber, which is preferably one of a highlynon-linear photonic crystal fiber, a side-hole fiber, a hollow-corephotonic bandgap fiber and a solid-core photonic bandgap fiber.

The optical pump laser may comprise a mode-locked laser. The pump pulserepetition rate may be variable. The optical pump laser mayalternatively comprise a modelocked fiber oscillator operable togenerate optical pump pulses. The optical pulse source preferablyfurther comprises an optical fiber amplifier configured to receive pumppulses from the modelocked fiber oscillator and operable to amplify thepump pulses. The modelocked fiber oscillator and the amplifier togetherform a master oscillator power amplifier (MOPA). The gating device maybe provided between the modelocked fiber oscillator and the opticalfiber amplifier.

The gating device preferably comprises an optical modulator element,such as an acousto-optic modulator, an electro-optic modulator or asemiconductor saturable absorber mirror (SESAM) modulator. The modulatorelement is preferably synchronised with the output of the optical pumplaser, to selectively control the repetition rate of pulses launchedinto the fiber, to thereby selectively control the output pulserepetition rate. The gating device is preferably operable to control theoutput pulse repetition rate from a few Hertz up to 100 MHz.

The optical pulse source preferably further comprises awavelength-tunable optical bandpass filter configured to receive anoptical supercontinuum pulse and to wavelength filter the pulse at apredetermined wavelength. The wavelength-tunable optical bandpass filteris preferably configured to receive a plurality of opticalsupercontinuum pulses in a pulse set and to wavelength filter each pulseat a respective one of a corresponding plurality of predeterminedwavelengths, to thereby form a wavelength-interleaved opticalsupercontinuum pulse set. The wavelength-tunable optical bandpass filteris preferably configured to filter a plurality of such pulse sets inseries to thereby form a repeating wavelength-interleaved opticalcontinuum pulse train.

The transmission spectrum of the wavelength-tunable optical filter canbe selected to pass any wavelength or set of wavelengths within thesupercontinuum spectrum, to thereby control the spectral profile of eachoutput optical pulse. The optical pulse source can thus be used as anoptical source for Pulse Interleave Excitation Forster Resonance EnergyTransfer (PIE-FRET).

The optical pulse source preferably further comprises second gatingdevice provided after the nonlinear optical element and configured topulse-pick received optical continuum pulses to a lower pulse repetitionrate. The second gating device preferably comprises an acousto-opticalmodulator or a wavelength-tunable optical bandpass filter. Thewavelength-tunable optical bandpass filter is preferably anacousto-optic tunable filter.

Preferably, a single wavelength-tunable optical bandpass filter isconfigured to perform both wavelength filtering and pulse-picking. Thesecond gating device is preferably further operable to reduce theoptical intensity of one or more optical pulses in the pulse set ortrain. The second gating device can thereby be operated to control theoptical intensity of the pulses in a train of optical pulses ofdifferent wavelengths, so that the optical intensity can be changed forpulses of different wavelengths.

The optical pulse source may alternatively further comprise: an opticalbeam splitter coupled to the output of the optical supercontinuum pulsesource, to thereby split the optical supercontinuum pulses into firstand second split pulses in first and second optical paths; first andsecond optical filters having different wavelength transmission spectrarespectively provided in the first and second optical paths; an opticaldelay element provided in the first optical path; and an optical beamcombiner arranged to recombine the first and second split pulses fromthe first and second optical paths into pulse train in a common opticalpath. A pulse train comprising interleaved optical supercontinuum pulsesof two different wavelengths is thereby produced.

According to a second aspect of the present invention there is provideda method of scaling the energy of optical supercontinuum pulsesgenerated using the optical pulse source as described above. The methodcomprises the steps of: reducing the pump pulse repetition rate of theoptical pump laser; and increasing the pump pulse duration. Thisinvention therefore enables the development of higher-energysupercontinuum pulse sources for certain applications where higher pulseintensity is required, with STED microscopy being one example.

The method may comprise reducing the pump pulse repetition rate to lessthan 20 MHz and increasing the pump pulse duration to greater than 10ps, and most preferably to at least 200 ps.

According to a third aspect of the present invention there is providedan illumination source for stimulated emission depletion microscopy. Theillumination source comprises an optical pulse source as describedabove, configured to generate output optical pulses having a repetitionrate in the range 1 MHz to 10 MHz and an energy spectral density of atleast 100 picojoules per nanometer (pJ/nm), the optical pulse sourcebeing operated to scale the energy of the optical supercontinuum pulsesby reducing the pump pulse repetition rate of the optical pump laser andincreasing the pump pulse duration.

The optical pulse source may be operated to reduce the pump pulserepetition rate to less than 20 MHz. The optical pump laser ispreferably operable to generate optical pump pulses having a pump pulseduration of greater than 10 ps, and most preferably at least 200 ps.

According to a fourth aspect of the present invention there is providedan optical fluorescence microscope based on stimulated emission anddepletion. The microscope comprises an illumination source as describedabove, with the illumination source being configured to provide bothexcitation illumination and stimulated emission illumination.

In conventional STED microscopes (see, e.g., G. Donnert et al.,Proceedings of the Natural Academic Society U.S.A 103, 11440-11445(2006)) two lasers are used simultaneously, one (typically a Ti:Sapphirelaser) with high optical intensity to form the STED beam and a secondtunable visible laser to excite the fluorescence. Conventionalsupercontinuum lasers, based on modelocked fiber laser-pumped PCF's orTi:Sapphire laser-pumped PCF's, are too low in pulse energy for STED.Using a reduced repetition rate and longer pulse according to thepresent invention, an optical pulse source is provided which is operableto generate supercontinuum pulses having a pulse energy spectral densityapproaching 1 nJ/nm, making this a suitable single-source solution forboth fluorescence excitation and STED within a STED microscope.

Another benefit of this invention is that it provides the ability to beflexible in pump pulse duration and the length of microstructure opticalfiber required to generate the supercontinuum output pulses.

The cases described within this invention relate specifically, but notexclusively to, the wavelengths falling within the gain bandwidth ofYtterbium doped silica fibers—between 1020 nm and 1080 nm. Equally, thisinvention can relate to supercontinuum pulses generated using a Nd-dopedfiber laser or Er-doped fiber laser or DPSS laser source as the opticalpump laser.

BRIEF DESCRIPTION OF THE DRAWINGS

The preferred embodiments of the present invention will now be describedby way of example only and with reference to the accompanying drawingsin which:

FIG. 1 is a schematic illustration of a prior art arrangement for thegeneration of an optical supercontinuum;

FIG. 2 is schematic illustration of a prior art supercontinuum fiberlaser;

FIG. 3 is a schematic illustration of an optical pulse source accordingto a first embodiment of the present invention generated from afree-space DPSS mode-locked laser source;

FIG. 4 is a schematic illustration of an optical pulse source accordingto a second, embodiment of the present invention;

FIG. 5 is a schematic illustration of an optical pulse source accordingto a third embodiment of the present invention;

FIG. 6 is a schematic illustration of an optical pulse source accordingto a fourth embodiment of the present invention;

FIG. 7A and FIG. 7B are representations of a first set of output opticalpulse trains exiting the microstructured optical fiber (FIG. 7A) andexiting the AOTF (FIG. 7B), which may be generated by the optical pulsesource of FIG. 6;

FIG. 8A and FIG. 8B are representations of a second set of outputoptical pulse trains exiting the microstructured optical fiber (FIG. 8A)and exiting the AOTF (FIG. 8B), which may be generated by the opticalpulse source of FIG. 6; and

FIG. 9A and FIG. 9B are representations of a third set of output opticalpulse trains exiting the microstructured optical fiber (FIG. 9A) andexiting the AOTF (FIG. 9B), which may be generated by the optical pulsesource of FIG. 6.

DETAILED DESCRIPTION

Referring to FIG. 3, a first embodiment of the invention provides anoptical pulse source 30 comprising an optical pump laser source 31, amicrostructured optical fiber 35 and a gating device 33 (also referredto below as “gating means” 33).

The optical pump laser 31 in this example takes the form of a diodepumped solid state (DPSS) laser source (for example a mode-locked 1064nm laser operating at 76 MHz), which is operable to produce pulses ofapproximately 200 nJ energy and duration 10 ps. The output pulses fromthe pump laser 31 pass through lens (e.g., a set of lenses) 32 to thegating means 33, which here comprises an optical modulator, synchronisedto the pump laser 31 and operable so that it can gate the pump pulsesand produce a pulse train having a reduced repetition rate (Rr), whereRr=Rf/N, where Rf is the pump laser fundamental frequency (in thisexample 76 MHz) and N is a positive integer. The pulses output from themodulator gating device 33 are injected via optics 34 into themicrostructured fiber 35, which here comprises a nonlinear element suchas a photonic crystal fiber (PCF). Propagation through the PCF 35 causesthe pulses to broaden spectrally to produce supercontinuum pulses at thereduced, output pulse, repetition rate Rr. FIG. 3 thus illustrates a lowrepetition rate picosecond or femtosecond supercontinuum pulse source,in this case pumped from a free-space DPSS mode-locked laser source.

One of the major drawbacks with the use of free-space optics, as shownin FIG. 3, is that launch stability into the small structures ofnonlinear PCF's is poor and the end face of these fibers can easily bedamaged. FIG. 4 shows an optical supercontinuum pulse source 40according to a second, all fiber embodiment of the invention, similar tothe arrangement shown in FIG. 2. The optical pump laser 41 comprises inthis example a mode-locked fiber oscillator operable at a repetitionrate Rf of 40 MHz to produce low energy pulses at 1064 nm and having apulse duration of approximately 10 ps. The pump pulses are amplified inan optical fiber amplifier 42. The pump pulses are then injected intogating device 43, which in this example is in the form of afiber-coupled modulator, which in this example comprises anacousto-optic modulator (AOM). It will be appreciated by one skilled inthe art that an Electro-Optic Modulator (EOM), SESAM modulator or anyother type of fast modulation device, capable of switching with risetimes of less than one microsecond may alternatively be used. Themodulator gating means 43 is synchronized to the oscillator frequency(Rf) and is operable to gate the oscillator output, i.e. the pumppulses, down to a repetition rate of a few tens of KHz, and hencepulse-pick the pump pulses, thereby providing a selectable pump pulsefrequency Rr=Rf/N (N is a positive integer value).

The reduced repetition rate pump pulses outputted from the modulatorgating means 43 are amplified in a power amplifier 45. The pump laser 41and the power amplifier 45 together form a master oscillator poweramplifier (MOPA) 46. The pump pulses output from the MOPA 46 have amaximum pulse peak power, governed by the nonlinear limitations of thepower amplifier 45 of, for example, approximately 40 KW, correspondingto a pump pulse energy of approximately 400 nJ.

Following amplification, the pump pulses are input into a length ofhighly nonlinear microstructured optical fiber 44, in this example PCF,with a zero dispersion wavelength at approximately 1040 nm. Propagationthrough the PCF 44 produces extensive spectral broadening of the pumppulses into the visible and further out into the IR region of thespectrum, thereby forming supercontinuum pulses. The supercontinuumpulses have a supercontinuum spanning from below 450 nm to greater than2000 nm, a total supercontinuum pulse energy of approximately 100 nJ,and energy spectral density of approximately 50 picoJoules/nm (pJ/nm).

In this example, since the oscillator remains the same, for a givenoutput pulse energy one achieves nominally identical spectra for allrepetition rates. The pulse energy is fixed and clamped, but the pulseto pulse separation increased with reducing repetition rate.

In an alternative arrangement, the optical pulse source 40 of thisembodiment may be operated with the low-power mode-locked fiberoscillator (pump laser) 41 generating pulses having a pulse duration of200 ps. Amplification within a fiber amplifier with nonlinear clamp onthe peak power of approximately 40 KW, results in a correspondingmaximum pump pulse energy of 8 μJ (for example, 1 MHz, 8 W average powerand 200 ps pulse duration). Supercontinuum pulses having a totalsupercontinuum pulse energy of approximately 2 μJ and energy spectraldensity of approximately 1 nJ/nm are produced.

Referring to FIG. 5, a third embodiment of the invention provides anoptical pulse source 50 which adds to the optical pulse source 40according to the second embodiment. In this embodiment, the opticalpulse source 50 further comprises a polarizing beam splitter 52 (anon-polarizing beam splitter or other similar splitting mechanism mayalternatively be used), optical filters 53, an optical delay element 54,and a beam combiner 56. The output of the optical pulse source 40 (seealso leftmost inset pulse plot) is passed through optics 51 and isthereafter split into two beam paths (A, B) by the beam splitter 52. Theoptical filters 53 are provided in each of the split beam paths. Theoptical filters 53 may be acousto-optic tunable filters (AOTF), eachbeing operable to transmit a different wavelength, or standard filtersof different transmission wavelengths.

The optical delay element 54, which takes the form of an optical fiberdelay line, is provided in one beam path (A). For a 20 MHz opticalsupercontinuum pulse repetition rate output from the optical pulsesource 40, a delay of 25 ns is required for the pulses travelling alongpath A. The optical fiber delay line 54 thus comprises a delay path ofbetween 5 m and 7.5 m in length. A coupling lens 55 is provided forlaunching the split pulses into the delay line 54. Propagation alongpath A causes the split optical pulses in that path to be delayed withrespect to the optical pulses in the other split beam path (B). It willbe appreciated that this can alternatively be achieved in free-space.

The beam combiner 56, which as shown comprises a dichroic mirror but mayalternatively comprise a fiber-coupler, is provided at the ends of thesplit beam paths A and B to recombine the two split pulse trains, tothereby form a pulse train comprising interleaved pulses of twodifferent wavelengths, as represented by the white and black arrows (seealso rightmost inset pulse plot). Although an optical fiber delay line54 is the preferred choice, this requires launching of the light into anoptical fiber 54 by a suitable lens 55, incurring additional loss andinstability. This approach is attractive as an experiment, but is lesspractical for a commercial instrument. In addition, this approach cannotuse a variable supercontiuum pulse repetition rate, since the requiredpath imbalance, introduced by the delay line, changes with repetitionrate.

A fourth embodiment of the invention provides an optical pulse source60, as shown in FIG. 6, comprising an optical pulse source 40 accordingto the second embodiment and a wavelength-tunable optical bandpassfilter 63. The optical pulse source 40 is operable to generate opticalsupercontinuum pulses having wavelength spectra from 400 nm to 2000 nm,at pulse repetition rates, variable between 40 MHz and less than 100kHz. The optical pulse source 40 is provided with an on-boardmicro-processor controller, which determines when the supercontinuumpulses are delivered, and provides a transistor-transistor logic (TTL)output trigger signal. The optical output of the optical pulse source 40is in free space, and the beam is collimated or waisted by optics 62.

In an example embodiment, the wavelength-tunable optical bandpass filter63 comprises an acousto-optic tunable filter (AOTF) driven by an RFdriver or Direct Digital Synthesiser (DDS) 64. The AOTF 63 is operableover a spectral range (for example from 400 nm to 700 nm) covered by theoptical supercontinuum pulse spectrum. The DDS 63 is synchronised withthe optical pulse source 40 either through an external trigger 61, orfrom a microprocessor, such that the AOTF can be modulated at theoperating frequency of the optical pulse source 40 or at any lowerfrequency. The AOTF 63 can also be operated in burst mode, to transmit aburst of optical pulses.

By changing the RF frequency of the AOTF driver 64, one can tune thefilter output to any wavelength lying within the AOTF 63 andsupercontinuum spectral ranges. Typical AOTF's have spectral ranges of,for example, from 400 nm to 700 nm in the visible region, from 700 nm to1100 nm in the near infra-red (NIR) and from 1100 nm to 2000 nm in theIR. All of these wavelengths are available within the spectrum of thesupercontinuum optical pulses generated by the optical pulse source 40.

The first order diffracted output of the AOTF 63 forms the output of theoptical pulse source 60, and may be launched into an optical fiber orused in free space. The zero-order of the AOTF, is either absorbed by asuitable beam dump 65 or can form a second output of the optical pulsesource 60. In this example, the AOTF 63 is triggered to operate at oneuser-defined wavelength for one pulse and a second user definedwavelength for the next pulse, thus generating a wavelength-interleavedoutput pulse train of alternating wavelength pulses, as shown in therightmost inset pulse plot. The optical pulse source 60 is thereforesuitable for use as an illumination source for PIE-FRET.

Dependent on the speed of the AOTF 63 (which is often limited by thespot size of the beam through the crystal), the AOTF can be configuredto deliver pulses at the optical supercontinuum pulse repetition rate,as shown in FIG. 7A and FIG. 7B, where FIG. 7A shows the pulses from theoptical pulse source 40 and FIG. 7B shows the AOTF output, i.e. theoutput of the optical pulse source 60, which comprises awavelength-interleaved train of filtered pulses at two differentwavelengths (represented by the solid and dashed lines).

The AOTF 63 may additionally be operable as a pulse-picker (i.e., agating device) to reduce the pulse repetition rate for the opticalsupercontinuum output pulses. As shown in FIG. 8A and FIG. 8B, thesupercontinuum pulse train is shown in FIG. 8A, and the AOTF outputpulse train of wavelength filtered and reduced repetition rate pulses attwo different wavelengths (represented by the solid and dashed lines) isshown in FIG. 8B.

The AOTF 63 is operable to filter the supercontinuum optical pulses atmore than two wavelengths, to produce a very complex pulse train whichmight have numerous applications within advanced imaging applications.FIG. 9A and FIG. 9B, for example, show the supercontinuum optical pulses(FIG. 9A) at a given repetition rate, and an output optical pulse trainfrom the optical pulse source 60 (FIG. 9B) with the AOTF 63 operating tofilter each set of four supercontinuum pulses at four differentwavelengths, to thereby generate an output optical pulse traincomprising interleaved optical pulses of four different wavelengths, asrepresented by the four different line types in the plot.

The AOTF 63 is also operable to amplitude control the output pulseintensities, which provides another degree of flexibility in optimisingsystem performance. This could, for example, balance the illuminationintensities for optical pulses at different wavelengths topre-compensate for different fluorescence levels from donor and acceptorfluorophores in FRET.

The combination of the variable optical supercontinuum pulse repetitionrate and AOTF to form a variable repetition rate and wavelength opticalpulse source, means that one can produce a series of repetition ratesand with arbitrary, wavelength varying pulse trains. It will beappreciated that the described variable repetition rate optical pulsesource may be replaced by a fixed repetition rate optical pulse sourcegenerating supercontinuum pulses, provided that the AOTF can operatesufficiently fast. For example, it is difficult to produce modelockedlasers at repetition rates much below 20 MHz, and this would require aswitching speed of the AOTF to be of the order of 10 ns, to reliablyswitch between colours with good extinction ratio between adjacentpulses—essential for FRET applications where very sensitive electronicsand detectors are used.

Typically, switching speeds of the 100 ns to microsecond level areachievable with AOTF's, meaning that an optical pulse repetition rate of5 MHz and below becomes a realistic solution. The variable repetitionrate optical pulse source presents a means to achieving this speed, andalso provides the flexibility to further vary the repetition frequency.

An example in which the optical pulse source 60 may be used as anillumination source, is in an imaging application known aspulsed-interleaved-excitation, most commonly used within FRET (ForsterResonance Energy Transfer). FRET is the radiation-less energy transferfrom an optically excited donor fluorophore to a nearby acceptormolecule through dipole-dipole interaction, and enables quantitativelength measurement due to the strong distance dependence of FRET. Inconventional FRET, the quality of measurement is hampered by incompleteFRET pairs (having a missing or non-functional acceptor molecule), whichlook very similar to FRET pairs with large donor-acceptor distances. Inaddition, determination of the FRET efficiency requires quantitativeinformation on the detection efficiency for donor and acceptorfluorescence—including spectral crosstalk.

Pulse interleave excitation FRET (PIE-FRET) overcomes these issues byhaving consecutive pulses operating at different wavelengths. The firstpulse operates at a wavelength to excite a donor molecule and initiateFRET. This leads to donor-to-acceptor fluorescence. The second pulseoperates at a wavelength to excite directly the acceptor fluorophore butnot the donor. This process determines whether or not the FRET pair hasa properly functioning acceptor. PIE-FRET has been demonstrated usingtwo different colour laser diodes, modulated at the same frequency butwith one diode delayed in time with respect to the other, producing apulse train of alternating colour pulses. Typically, this process iscarried out at fairly high repetition rates (20 MHz, to 40 MHz) withpulse-to-pulse time separations of the order of 50 ns to 25 nsrespectively, which is usually acceptable for measurement of mostbiological lifetimes, and provides high throughput for building up largeimages. The restriction of the two diode approach is that thewavelengths are limited to those available from suitable laser diodes.The optical pulse source 60 of the present invention allows theproduction of pulse trains having a combination of any sets ofwavelengths within the supercontinuum spectral range.

In a fifth embodiment of the invention, the use of a fiber laserincorporating a pulse-picker element allows for the delivery of higherenergy supercontinuum lasers within the visible region of the spectrum.

In some examples, it is important to be able to scale the energy fromthe supercontinuum optical pulses—particularly within the visible regionof the spectrum. Using the same system configuration as FIG. 4, it ispossible to scale the average supercontinuum energy by reducing the pumppulse repetition rate but increasing the pump pulse duration to hundredsof picoseconds. This effectively results in a higher energy 1064 nmfundamental input pulse (the peak power of the pulse is limited withinthe fiber amplifier by nonlinear interactions dominated by stimulatedRaman scattering (SRS)) and a corresponding increase in the averageoutput supercontinuum pulse energy across the supercontinuum spectraexiting the PCF.

By increasing the pulse duration, one therefore increases the averagepulse energy attainable within the SRS limit of the fiber amplifier. Theresult, after injection into the PCF, is an increase in the resultingspectral energy density of the supercontinuum pulses. For example, astandard supercontinuum laser operating at 20 MHz repetition rate anddelivering 2 W average power in the supercontinuum, produces asupercontinuum pulse energy of approximately 100 nJ. When the visibleregion of this source output is filtered, the average spectral energydensity is in the region of 50 pJ/nm (400 mW over a 400 nm bandwidth).

In known commercial supercontinuum fiber lasers, such as theaforementioned SC450 from Fianium Ltd., or the SuperK from Koheras A/S,the average energy spectral density of pulses within the visible regionof the spectrum ranges from approximately 35 pJ/nm (SuperK Rapid, 3mW/nm at 80 MHz) to 50 pJ/nm (SC450, 1 mW/nm at 20 MHz). For thesesources, the total supercontinuum power ranges from 2 W to 6 W and isgenerated using pulsed optical pump sources of between 8 W and 20 Waverage power. The spectral broadening in the photonic crystal fiber isgenerated by the high peak power of the pump source optical pulses whichcan be as high as 40 KW (8 W, 20 MHz, 10 ps). As an example, a typicalSC450-2 supercontinuum laser from Fianium Ltd., with a power spectraldensity in the visible of 1 mW/nm, has an energy spectral density ofapproximately 50 pJ/nm (50×10⁻¹² J/nm), and is pumped by a pulsedoptical pump source operating at 20 MHz with a pulse duration ofapproximately 10 ps and an output power of approximately 8 W.

In order to scale the energy spectral density of these knowsupercontinuum sources at these repetition rates to 500 pJ/nm, pumplaser source having between 10 and 15 times the output power would berequired, equating to a pulse optical pump source with average power ofbetween 80 W and 285 W.

In this embodiment, a pulse-picked optical pulse source 40, as shown inFIG. 6, operating at a 5 MHz pulse repetition rate and a 10 m length ofPCF 44, achieves the same visible power and spectral bandwidth but thisresults in a 5-Fold increase in spectral energy to 200 pJ/nm Startingwith a 200 ps optical pulse and a standard length of nonlinear fiber, itis possible to scale the spectral energy density to in excess of 1 nJ/nmat 1 MHz, with an average visible power of approximately 400 mW (over a400 nm band) at 1 MHz.

A sixth embodiment of the invention comprises a pulse-picked opticalpulse source 40, as shown in FIG. 6, in which the pump pulse repetitionrate is reduced to 1 MHz, and the pulse duration increased toapproximately 200 ps. In order to generate the required spectralbroadening within the PCF 44, 40 KW of pulse peak power is attained fromthe pulsed pump source 41 at an average output power of 8 W, resultingin a 2 W supercontinuum, with 1 mW/nm power spectral density in thevisible spectral region, corresponding to approximately 1 nJ/nm energyspectral density.

A seventh embodiment of the invention provides a method of scaling theenergy of optical supercontinuum pulses generated using the opticalpulse sources 30 or 40 of the previously described example embodiments.The method comprises reducing the pump pulse repetition rate of theoptical pump laser 31 or 41 and increasing the pump pulse duration.

In general, the magnitude of spectral broadening of an optical sourcepropagating through a given nonlinear optical element is directlyproportional to the intensity of that source. For a pulsed laser source,the intensity is given by the pulse peak power:P _(peak) =P _(avg)/(R*dT)where P_(peak) is the peak power of the pulse, P_(avg) is the averagepower of the laser source, R is the pulse repetition frequency of thesource and dT is the temporal duration of the pulse. The energy of thepulse is given by:E=P _(avg) /R

For a laser based on a MOPA (Master Oscillator, Power Amplifier) design,while the average power of the laser can be scaled, the peak power islimited by nonlinear effects (mainly stimulated Raman scattering) withinthe fiber amplifier system, which has the effect of clamping the peakpower, resulting in:P _(peak)=Constant=P _(avg)/(R*dT)

The method realises that, to a first approximation, the extent ofspectral broadening within a pulsed supercontinuum fiber laser can beconsidered to be limited by the peak power of the pulse, but that theenergy of the pulse can be changed by adjusting the pulse repetitionrate and the temporal duration of the pulse.

The maximum pulse energy of the optical pulse source 30 and 40, andhence the maximum pulse energy (or energy spectral density) of resultingsupercontinuum pulses, can therefore be increased by reducing the pulserepetition rate of the pump laser 31 and 46 and, in the case of the MOPA46 by either reducing the pulse repetition rate or scaling the averagepower from the amplifier 45. For a given amplifier design and averagepower output, an increase in pulse energy can be achieved with areduction of pump pulse repetition rate and increase of pump pulseduration.

The limit of peak power extractable from a given fiber amplifier isdetermined by the length of the amplifier fiber and the cross sectionalarea of the mode propagating within this fiber. For standard single modeoptical fibers operating at 1 μm, the peak power limit can be as low asa few kW, but this can be scaled to tens of kW for large mode area (LMA)fibers where the effective mode field diameter within the fiber mode canbe greater than 12 μm.

The described embodiments make use of the reduction in pulse repetitionrate to enable scaling of the energy spectral density of thesupercontinuum without the requirement for scaling the power spectraldensity and therefore the average power from the pulsed optical pumpsource.

From the foregoing therefore, it is evident that the present inventionenables the delivery of supercontinuum spectra or filtered visible lasersources at a range of pulse repetition rates from 100 MHz down to a fewHz or less. This invention has clear benefits over conventionalsupercontinuum sources for applications where the pulse-to-pulseseparation of the source is an important parameter.

Various modifications may be made without departing from the scope ofthe present invention. For example, where the DPSS pump laser may bereplaced by a different type of mode-locked laser, and may comprise aMOPA. The PCF may be replaced by a different micro-structured opticalfiber, such as a side-hole fiber, hollow-core photonic bandgap fiber orsolid-core photonic bandgap fiber, or may be replaced by a differentnonlinear optical element, such as a nonlinear crystal like such as LBO,BBO, CLBO, PPLN, PPKTP or any other nonlinear optical crystal phasematched to produce frequency conversion of the pump laser output. Also,in the embodiments of the invention comprising a MOPA, the gating devicemay alternatively be located after the power amplifier

The invention claimed is:
 1. Variable repetition rate and wavelengthoptical pulse source, comprising: a variable repetition rate source ofsupercontinuum pulses, said variable repetition rate source ofsupercontinuum pulses comprising a nonlinear optical element comprisingan optical fiber arranged to receive pump pulses and to spectrallybroaden the pump pulses to generate the supercontinuum pulses; saidvariable repetition rate source of supercontinuum pulses providingnominally identical spectra for different repetition rates; and awavelength tunable optical bandpass filter to filter the supercontinuumpulses, wherein said source of supercontinuum pulses and said wavelengthtunable optical bandpass filter are configured such that the opticalpulse source can provide variable repetition rate and variablewavelength optical pulses.
 2. The variable repetition rate andwavelength optical pulse source of claim 1 wherein the wavelengthtunable optical bandpass filter is configured to receive a plurality ofsupercontinuum pulses in a pulse set and to wavelength filter each pulseat a respective one of a corresponding plurality of predeterminedwavelengths, to thereby form a wavelength interleaved optical pulse set.3. The variable repetition rate and wavelength pulse source of claim 1wherein the wavelength tunable optical bandpass filter is configured tooperate in burst mode to provide a pulse train comprising a burst ofoptical pulses.
 4. The variable repetition rate and wavelength opticalpulse source of claim 1 wherein the wavelength tunable optical bandpassfilter comprises an acousto-optic tunable filter (AOTF).
 5. Variablerepetition rate and wavelength optical pulse source, comprising: avariable repetition rate source of supercontinuum pulses comprising anonlinear optical element comprising an optical fiber arranged toreceive pump pulses and to spectrally broaden the pump pulses togenerate the supercontinuum pulses; a pump pulse source that can providethe pump pulses having different repetition rates, wherein the pulseenergy is fixed as pulse to pulse separation is increased with reducingrepetition rate; and a wavelength tunable optical bandpass filter tofilter the supercontinuum pulses, wherein said source of supercontinuumpulses and said wavelength tunable optical bandpass filter areconfigured such that the optical pulse source can provide variablerepetition rate and variable wavelength optical pulses.
 6. A method ofproducing variable repetition rate and variable wavelength opticalpulses, comprising: providing supercontinuum pulses using a variablerepetition rate supercontinuum pulse source, including spectrallybroadening pump pulses propagating in an optical fiber; reducing therepetition rate of the pump pulses, including fixing the pulse energy asthe pulse to pulse separation is increased, to thereby providesupercontinuum pulses having a reduced repetition rate; and filteringthe supercontinuum pulses using a wavelength tunable optical bandpassfilter, thereby providing optical pulses having different repetitionrates and different wavelengths.
 7. Variable repetition rate andwavelength optical pulse source, comprising: a variable repetition ratesource of supercontinuum pulses; a wavelength tunable optical bandpassfilter to filter the supercontinuum pulses at two or more wavelengths,wherein said source of supercontinuum pulses and said wavelength tunableoptical bandpass filter are configured such that the wavelength tunableoptical filter can be modulated, synchronously with the source ofsupercontinuum pulses, at the operating frequency of the source ofsupercontinuum pulses or at a lower frequency, wherein the optical pulsesource can provide variable repetition rate and variable wavelengthoptical pulses including a series of repetition rates with selectedwavelength-varying pulse trains; and wherein the source ofsupercontinuum pulses comprises an optical pump laser operable togenerate optical pump pulses at a pump pulse repetition rate; anonlinear optical element; a gating device, provided between the pumplaser output and the nonlinear optical element, and arranged toselectively control the launch of pump pulses into the nonlinear opticalelement at a variable, lower repetition rate, to thereby selectivelycontrol the repetition rate of optical supercontinuum pulses generatedwithin the nonlinear optical element.
 8. The variable repetition rateand wavelength optical pulse source of claim 7 wherein the nonlinearoptical element comprises a microstructured optical fiber.
 9. Thevariable repetition rate and wavelength optical pulse source accordingto claim 7 wherein the optical pump laser comprises a fiber oscillatorand wherein the source of supercontinuum pulses comprises an opticalfiber amplifier configured to receive pump pulses from the modelockedfiber oscillator and operable to amplify the pump pulses, the opticalfiber amplifier being provided between the gating device and the fiberoscillator.
 10. The variable repetition rate and wavelength opticalpulse source according to claim 8 wherein the optical pump lasercomprises a fiber oscillator and wherein the source of supercontinuumpulses comprises an optical fiber amplifier configured to receive pumppulses from the modelocked fiber oscillator and operable to amplify thepump pulses, the optical fiber amplifier being provided between thegating device and the fiber oscillator.
 11. The variable repetition rateand wavelength optical pulse source of claim 9 wherein the source ofsupercontinuum pulses comprises a further optical amplifier providedbetween the gating device and the nonlinear optical element.
 12. Thevariable repetition rate and wavelength optical pulse source accordingto claim 10 wherein the source of supercontinuum pulses comprises afurther optical amplifier provided between the gating device and thenonlinear optical element.
 13. The optical pulse source according toclaim 1 configured as a pulsed interleaved excitation apparatusproviding first and second interleaved pulses, the first pulse having awavelength selected to excite a donor molecule and the second pulsehaving a different wavelength selected to excite an acceptor molecule,the pulse interleaved excitation apparatus comprising said optical pulsesource as an illumination source for providing the first and secondpulses having different wavelengths.
 14. A method of producing variablerepetition rate, wavelength-varying optical pulses, comprising:providing supercontinuum pulses using a variable repetition ratesupercontinuum pulse source, including spectrally broadening pump pulsespropagating in an optical fiber; reducing the repetition rate of thesupercontinuum pulses to provide supercontinuum pulses having a reducedrepetition rate, including providing nominally identical spectra fordifferent repetition rate supercontinuum pulses; and filtering thesupercontinuum pulses at two or more wavelengths using a wavelengthtunable optical bandpass filter, thereby providing optical pulses havingdifferent repetition rates and different wavelengths.
 15. The method ofclaim 14, wherein filtering the supercontinuum pulses at two or morewavelengths comprises: receiving, at the wavelength tunable opticalbandpass filter, a plurality of supercontinuum pulses in a pulse set;and wavelength filtering each pulse at a respective one of acorresponding plurality of predetermined wavelengths, to thereby form awavelength interleaved optical pulse set.
 16. The method of claim 14,comprising providing a pulse train comprising a burst of optical pulsesby operating the wavelength tunable optical bandpass filter in a burstmode.
 17. The method of claim 14 comprising launching the pump pulsesinto the optical fiber without the use of free space optics. 18.Variable repetition rate and wavelength optical pulse source,comprising: a fixed or variable repetition rate source of supercontinuumpulses; a wavelength tunable optical bandpass filter to filter thesupercontinuum pulses at two or more wavelengths, wherein said source ofsupercontinuum pulses and said wavelength tunable optical bandpassfilter are configured such that the optical pulse source can providevariable repetition rate and variable wavelength optical pulsesincluding a series of repetition rates with selected wavelength-varyingpulse trains, including providing one wavelength for one pulse and asecond wavelength for the next pulse, wherein said optical pulse sourcecan form a wavelength interleaved optical pulse train.
 19. The variablerepetition rate and wavelength optical pulse source of claim 1 whereinthe pump pulses are launched into the optical fiber without the use offree space optics.
 20. The variable repetition rate and wavelengthoptical pulse source of claim 18 wherein said fixed or variablerepetition rate source of supercontinuum pulses comprises a fixedrepetition rate source of supercontinuum pulses.
 21. The variablerepetition rate and wavelength optical pulse source of claim 18 whereinsaid fixed or variable repetition rate source of supercontinuum pulsescomprises a variable repetition rate source.
 22. The variable repetitionrate and wavelength optical pulse source of claim 18 wherein said fixedor variable repetition rate source of supercontinuum pulses comprises amode locked laser.
 23. Variable repetition rate and wavelength opticalpulse source for providing output optical pulses, comprising: a fixed orvariable repetition rate source of supercontinuum pulses; a wavelengthtunable optical bandpass filter to filter the supercontinuum pulses attwo or more wavelengths, wherein said source of supercontinuum pulsesand said wavelength tunable optical bandpass filter are configured suchthat the optical pulse source can provide variable repetition rate andvariable wavelength optical pulses including a series of repetitionrates with selected wavelength-varying pulse trains, including selectivecontrol of the spectral profile of each output optical pulse.
 24. Thevariable repetition rate and wavelength optical pulse source of claim 1wherein said fixed or variable repetition rate source of supercontinuumpulses comprises a mode locked laser.
 25. The variable repetition rateand wavelength optical pulse source of claim 5 wherein the pump pulsesare launched into the optical fiber without the use of free spaceoptics.
 26. The method of claim 6 further comprising launching the pumppulses into the optical fiber without the use of free space optics.